Establishment of SRAP-PCR reaction system in apple
SI Peng
DAI Hong-yi
XUE Hua-bo
ZHANG Yu-gang
GUO Jun-ying
CAO Shang-yin
Abstract：Experiment was conducted with the apple cultivar Telamon and the F1 seedlings of Telamon × Fuji. The genomic DNA was extracted from the leaves by modified CTAB method. Orthogonal design was applied to optimize SRAP-PCR amplification system of apple in 5 factors such as Mg~(2+),dNTPs,Primer,Taq DNA polymerase and template DNA at 4 levels. The result of PCR was analyzed by intuitive analysis and variance analysis. An optimal reaction system(10 μL) was established, i.e.1 μL 10×buffer (Mg~(2+) free),2.0 mmol·L~(-1) Mg~(2+),0.8 mmol·L~(-1) dNTPs,0.2 mmol·L~(-1) Primer,0.6 U Taq DNA polymerase and 60 ng template DNA.48 F1 of Telamon × Fuji and other SRAP-PCR primer were used to test the stability of the optimized reaction system. The results indicated that the optimized reaction system was stable.
Keywords：AppleTelamonSRAP-PCROrthogonal analysis
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 168-173 )